Fig. 3: circATXN1 knockdown enhanced Hypoxia-Induced Partial EndMT in HCMECs.
From: Regulation of partial endothelial-to-mesenchymal transition by circATXN1 in ischemic diseases
a FUCCI images of HCMECs infected with Ad Scr-sh or Ad circATXN1-sh for 48 h and exposed to hypoxia for 24 h (red for G1, yellow for G1/S, green for S/G2-M) (n = 3 biological replicates, scale bars = 100 µm). b–c Transwell migration and spheroid sprouting assays of HCMECs infected with Ad Scr-sh or Ad circATXN1-sh, with hypoxia stimulation for 24 h (n = 4 biological replicates for b, scale bars = 100 µm. n = 6 biological replicates for c, scale bars = 100 µm). d The viability of HCMECs infected with Ad Scr-sh or Ad circATXN1-sh for 48 h and exposed to hypoxia for 24 h was determined by a CCK-8 assay (n = 8 biological replicates). e Tube formation assays of HCMECs infected with Ad Scr-sh or Ad circATXN1-sh for 48 h and exposed to hypoxia for 24 h (n = 4 biological replicates, scale bar, 100 μm). f Representative HE stained sections showed luminal structures containing erythrocytes in implants receiving HUVECs infected with Ad Scr-sh or Ad circATXN1-sh after 7 days. Scale bar = 100 μm. Quantification of red blood cells filled microvessel density as microvessels/mm2 in shown in the right panel. (n = 4 mice per group). g Immunofluorescence for CD31 (green) and αSMA (red) in HCMECs infected with Ad Scr-sh or Ad circATXN1-sh, with DAPI counterstaining (blue) (n = 8 IF images, scale bars = 20 μm). h Western blot analysis of Fibronectin, VE-cadherin, CD31, αSMA, and FSP1 in HCMECs infected with Ad Scr-sh or Ad circATXN1-sh, with β-actin as the internal control (n = 3 biological replicates). Statistical analysis: unpaired two-tailed Student t test for (b, c, d, e, f).
